Synthesis and fluorescence properties of substituted 7-aminocoumarin-3-carboxylate derivatives

4-Trifluoromethyl- or 6-bromo-substituted 7-diethylaminocoumarin-3-carboxamide derivatives 2 and 3, each containing a maleimide have been synthesized as potential fluorescent labeling reagents for thiol groups in proteins and their fluorescence properties have been determined. The 4-trifluoromethyl substituted compound 2 has a significantly greater Stokes shift than the comparable compound lacking this group, but both the new coumarins have low fluorescence quantum yields (phi (f)). When a 4-trifluoromethyl substituent is present, the 3-carboxamide is unusually labile to hydrolysis. Bromination of ethyl 7-diethylaminocoumarin-3-carboxylate 17 gave the 6- and 8-bromo derivatives 18 and 19 respectively, and also the 8-bromo-7-monoethylamino compound 20. phi (f) for the latter compound is 100-fold greater than for its diethylamino analogue 19, Fluorescence lifetime measurements support an interpretation based on the twisted intramolecular charge transfer (TICT) model to explain these large differences in phi (f).